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[Change of Ankyrin G Promoter Activity Following Treatment with Sodium Valproate at Different Concentrations]
Cui Liu1, Jie Wu1, Xiao-long Sui1
1Key Laboratory of Human Diseases Animal Models,State Administration of Traditional Chinese Medicine,Key Laboratory of Human Diseases Comparative Medicine of Ministry of Health,Institute of Medical Laboratory Animal Science,CAMS and PUMC,Beijing 100021,China.
Objective:
To investigate whether sodium valproate (VPA) directly regulates the activity of Ankyrin G(AnkG) promoter in vitro.
Methods:
The mouse AnkG promoter sequence was identified by comparing both human and mouse AnkG promoter sequences. The promoter was amplified from C57BL/6 mouse genome DNA and cloned into pGL3 Luciferase reporter vector. The Luciferase activity was detected in N2a and 293T cells and then treated with 0,0.5, and 1 mmol/L VPA for 12 h. The transcription activity of AnkG promoter in cells and the activity of VPA-treated Luciferase reporter vector in cells were detected using dual Luciferase reporter assay.
Results:
The AnkG promoter clone and its expression vector were successfully established, as confirmed by enzyme digestion and sequencing. The AnkG promoter showed high transcription activity in both N2a and 293T cells. The Luciferase activity was significantly induced following 0.5 mmol/L VPA treatment in both N2a and 293T cells. CONCLUSIONS VPA can up-regulate the AnkG expression via directly increasing its transcription activity. Thus, the in vivo AnkG expression may be directly regulated by the VPA at transcriptional level.
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